Cleaved parp asp214 d64e10 xp

Drug treated cells were washed in PBS and lysed in RIPA buffer (supplemented with protease and phosphatase inhibitors) (Sigma, St. Louis, MO) on ice for 30 minutes. Total protein concentrations were determined using a BCA kit (Thermo Scientific, Rockford, IL). Western blots were performed by resolving proteins by SDS-PAGE and transfer to nitrocellulose membranes. Membranes were probed with antibodies to: c-Myc (9E10) (sc-40); HRP conjugated GAPDH (FL-335) (sc-25778 HRP) (all from Santa Cruz Biotechnology); Max (AF4304) (R&D Systems); Cleaved PARP (Asp214) (D64E10) XP (5625; Cell Signaling). Proteins were visualized using horseradish peroxidase-conjugated secondary Mouse or Rabbit (1:5000) (GE Healthcare) or Goat (1:1000) (R&D Systems) antibodies and the SuperSignal ELISA Femto Maximum Sensitivity Substrate kit was used for detection (Thermo Scientific).

Imatinib mesylate, nilotinib, dasatinib, and CX-4945 were purchased from Santa Cruz Biotechnology, Inc. All inhibitors were dissolved as 10 mM stock solution in DMSO (Sigma-Aldrich Inc., Missouri, USA) and stored at -20°C. Protease inhibitor cocktails and Phosphatase inhibitor cocktails were purchased from Calbiochem and SIGMA-ALDRICH, respectively. Non-conjugated primary antibodies casein kinase IIα (D-10 and E-7), casein kinase IIα´ Antibody (E-7), casein kinase IIβ Antibody (6D5), and Actin (anti-actin antibody clone AC-15, SIGMA-ALDRICH) or beta-tubulin (β-Tubulin (9F3) were purchased from SantaCruz Biotechnology (CA, USA). Anti-HSP90 beta (phospho S226) antibody was purchased from Abcam (UK). Phospho-CK2 Substrate [(pS/pT)DXE] MultiMab™ Rabbit mAb, Phospho-CDC37 (Ser13) (D8P8F) Rabbit mAb, Phospho-CrkL (Tyr207) Rabbit mAb, Cleaved PARP (Asp214) (D64E10) XP, and HRP conjugated secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).